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目的:建立以硝苯地平为替代对照品同时测定银杏叶片中槲皮素、山柰素和异鼠李素含量的方法。方法:以硝苯地平为替代对照品，采用HPLC法，色谱柱为JADE-PAK十八烷基硅烷键合硅胶柱(4.6 mm×250 mm，5 μm)，流动相为0.5%磷酸溶液(用三乙胺调节pH2.2)-甲醇(50∶50)，硝苯地平检测波长为236 nm，槲皮素、山柰素和异鼠李素检测波长360 nm，柱温35 ℃，通过测定硝苯地平与槲皮素、山柰素和异鼠李素的相对校正因子，采用内标法，并利用相对校正因子计算银杏叶片中槲皮素、山柰素和异鼠李素含量。结果:硝苯地平、槲皮素、山柰素和异鼠李素进样量分别在0.12~1.16 μg、0.07~0.73 μg、0.06~0.56 μg、0.03~0.27 μg范围内与其峰面积线性关系良好；硝苯地平与槲皮素、山柰素和异鼠李素之间的相对校正因子分别为0.782 2、0.768 5、0.808 8；槲皮素、山柰素和异鼠李素平均回收率分别为98.48%、98.03%和94.55%。结论:用硝苯地平作为替代对照品，对银杏叶片中槲皮素、山柰素和异鼠李素含量进行测定和质量控制具有经济性、可行性和实用性。“,”Objective:To establish a method which Nifedipine was used as an alternative control for determinating of three flavonoid glycosides in the tablets of Ginkgo biloba leaves by substitution method of reference substance.Methods:Make Nifedipine instead reference substance, the analysis was performed on a JADE-PAK C18 column (4.6 mm × 250 mm, 5 μm), with the mobile phase consisted of 0.5% phosphoric acid solution-methanol (adjusting pH value to 2.2) (50:50) at the flow rate of 1.0 ml/min, the column temperature was 35 ℃. The Nifedipine was selected as the reference substance of three flavonoid glycosides and the correction factor of Nifedipine. The content of total flavonoid glycosides was determined by relative correction factor f and substitute reference.Results:The linear ranges of Nifedipine, quercetin, kaempferide and isorhamnetin were 0.17-1.73 μg, 0.11-1.08 μg, 0.08-0.84 μg, 0.04-0.40 μg. The relative correction factor ( n f) between Nifedipine and quercetin, kaempferide, isorhamnetin were 0.782 2, 0.768 5, 0.808 8, respectively. The average recovery (n n=6) were 98.48%, 98.03%, 94.55%.n Conclusion:The results indicated that the HPLC method by substitute reference substance is economical, feasible and practical.